Titanium dioxides (TiO2) film is one of the important biomaterials to improve biocompatibility of titanium (Ti). We have used a film coating method with an aerosol beam and femtosecond laser irradiation to form periodic structures on biomaterials for control of the cell spreading. The control of cell spreading on biomaterials is important for the development of advanced biomaterials. In this study, nanostructures with periods of 130 and 230 nm were formed on a film using a femtosecond laser with wavelengths of 388 and 775 nm, respectively. The nanostructure period on the film was 30% of the laser wavelengths. After cell test, cell spreading along the grooves of the periodic nanostructures was observed on the periodic nanostructures with the periods of 130 and 230 nm.
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